Tien-Shang Huang

Baicalein and baicalin are potent inhibitors of angiogenesis: Inhibition of endothelial cell proliferation, migration and differentiation

In recent studies, we have shown that baicalein and baicalin, 2 major flavonoids of Scutellaria baicalensis Georgi, exhibit anticancer activity against several cancers in vitro. In our present study, we assessed their potential as anti‐angiogenic agents in vivo employing chicken chorioallantoic membrane (CAM) assay and in vitro human umbilical vein endothelial cells (HUVECs) culture. When CAMs were treated with either baicalein or baicalin for 48 hr, the angiogenic response induced by basic fibroblast growth factor (bFGF) was markedly reduced in a dose‐dependent manner. Further characterization showed that both flavonoids exhibited dual antiproliferative (at low dose) and apoptogenic (at high dose) effects on HUVECs. In biochemical analysis, treatment of HUVECs with baicalein and baicalin for 24 hr resulted in a dose‐dependent decrease in the matrix metalloproteinase (MMP)‐2 activity. Moreover, the migration of endothelial cells and the differentiation of endothelial cells into branching networks of tubular structures in vitro were also inhibited by these 2 flavonoids in a dose‐dependent manner. Baicalein is more potent than baicalin in anti‐angiogenesis in vivo as well as in vitro. Taken together, the results of our study provide evidence that baicalein and baicalin possess an anti‐angiogenesis potential that is a previously unrecognized biologic activity.

Circulating levels of markers of inflammation and endothelial activation are increased in men with chronic spinal cord injury.

BACKGROUND/PURPOSE Accelerated atherogenesis is often seen in individuals with chronic spinal cord injury (SCI). However, the mechanisms contributing to this phenomenon remain unclear. This study aimed to evaluate whether SCI per se is associated with a low-grade chronic inflammatory state and endothelial activation, both of which are well-documented prerequisites for atherogenesis. METHODS Serum levels of markers of inflammation (C-reactive protein [CRP], interleukin-6, and soluble CD40 ligand) and endothelial activation (endothelin-1, soluble intercellular adhesion molecule-1, and soluble vascular cell adhesion molecule-1 [sVCAM-1]) were measured in SCI patients with CRP levels < 10 mg/L and with no evidence of active infection. Sixty-two men with traumatic neurologically complete SCI (20 tetraplegics and 42 paraplegics) and 29 age-matched male controls were enrolled. RESULTS Compared with able-bodied controls, subjects with SCI had a significantly lower body mass index (BMI) (-7%) and significantly lower serum levels of albumin (-10%), creatinine (-20%), low-density lipoprotein cholesterol (-10%), and high-density lipoprotein (HDL) cholesterol (-25%), and showed a trend toward higher fasting insulin levels. Irrespective of injury level and duration, subjects with SCI had significantly higher serum levels, compared to able-bodied controls, of CRP (mean, 4.0 +/- 2.7 mg/L vs. 1.4 +/- 1.1 mg/L), interleukin-6 (median, 2.5 pg/mL vs. 0.4 pg/mL; range, 1.5-3.6 pg/mL vs. 0.2-0.5 pg/mL), endothelin-1 (mean, 1.3 +/- 0.4 pg/mL vs. 0.9 +/- 0.3 pg/mL), and sVCAM-1 (mean, 1170 +/- 318 ng/mL vs. 542 +/- 318 ng/mL). The serum levels of all four factors correlated negatively with levels of serum albumin, creatinine and HDL cholesterol, but not with BMI or fasting insulin levels. In multivariate analyses, SCI was the only factor that was independently associated with increased serum levels of CRP, interleukin-6, endothelin-1 and sVCAM-1 after adjustment for confounding factors such as serum albumin and creatinine levels and parameters of dyslipidemia and insulin resistance. CONCLUSION In this study, we have, for the first time, demonstrated that SCI per se is associated with a low-grade chronic inflammatory state and endothelial activation, which may partly explain the increased atherogenic risk in patients with long-standing SCI.

Thyroid hormone changes cardiovascular surgery and clinical implications

Abstract Alterations in serum concentrations of total triiodothyronine (TT 3 ), total thyroxine (TT 4 ), and thyroid-stimulating hormone (TSH) frequently occur in patients with nonthyroidal illnesses. These changes correlate with the severity of the illness and the prognosis. In this study, 44 patients undergoing a cardiovascular operation had significant declines in serum TT 3 and TT 4 levels during cardiopulmonary bypass and thereafter. Serum TT 3 and TT 4 concentrations reached their nadir at 30 minutes after the start of cardiopulmonary bypass with values (mean ± standard error of the mean) of 0.77 ± 0.12 nmol/L (50.4 ± 7.6 ng/dL) and 68.2 ± 10.2 nmol/L (5.30 ± 0.79 μg/dL), respectively. The mean serum concentrations of TSH and TT 4 returned to preoperative levels by the sixth day after operation, whereas TT 3 levels remained low throughout the study period. The patients whose recovery was uneventful had higher serum TT 3 , TT 4 , and TSH levels than those who had complications or died. The trend toward recovery was initiated by a sharp increase in the serum TSH level and increases in serum TT 3 and TT 4 concentrations on the fourth day after operation. Patients with complications either did not show these changes or had only a transient increase in TT 3 and TT 4 levels. All of the patients had a normal serum free T 4 level before anesthesia. Those with an uneventful recovery had a higher serum free T 4 level on the sixth day after operation than those with complications. Two patients in the latter group had serum free T 4 levels less than normal at that time. The alterations in serum TT 4 , TT 3 , and TSH concentrations has no correlation with drugs (furosemide, dopamine hydrochloride, dobutamine hydrochloride, isoproterenol hydrochloride, norepinephrine, or epinephrine) administered during the study period. In summary, serum TT 3 , TT 4 and TSH levels declined in patients undergoing a cardiovascular operation, especially in those who had complications or died. The administration of catecholamines or furosemide was not an important factor in the development of abnormal thyroid function test results in our study.

Serum thyroid hormone binding inhibitor in nonthyroidal illnesses

We have employed the recently developed competitive ligand binding assay (CLBA) to study thyroid hormone binding inhibitor (THBI) in ether extracts of sera of 25 patients admitted to the Medical Intensive Care Unit with a variety of nonthyroidal illnesses (NTI). THBI was detected in 60% (15/25) of patients using one sample/patient and in 88% (15/17) using multiple (two to six) samples from different days. Mortality rate and mean serum concentrations of total T4, total T3, and albumin were similar in THBI-positive and THBI-negative patients. There was a tendency for a higher frequency of low serum total T4 in THBI-positive (10/15) than in THBI-negative (3/10) patients but the difference was not statistically significant (P less than 0.1 by Chi square). However, the mean dialyzable fraction of T4 (DFT4 0.11 +/- 0.02%, n = 9 v 0.054 +/- 0.004%, n = 10) and DFT3 (0.54 +/- 0.05% v 0.40 +/- 0.032%) were both significantly (P less than 0.05) higher in THBI-positive patients than THBI-negative patients. There was a significant correlation between THBI and DFT4 (r = 0.55, P less than 0.02) or DFT3 (r = 0.54, P less than 0.02). Prior extraction of serum with ether reduced DFT4 in NTI patients with high baseline DFT4 but not in normal subjects or NTI patients with mildly abnormal baseline DFT4. Addition to a normal serum (0.1 mL) of evaporated ether extract of a pooled NTI serum (0.10- to 3.0-mL equivalent) increased DFT4 progressively from 0.025% to 0.14%. Similar extract of a pooled serum of normal subjects had little or no effect.(ABSTRACT TRUNCATED AT 250 WORDS)

To evaluate the effectiveness of health care ethics consultation based on the goals of health care ethics consultation: a prospective cohort study with randomization

BackgroundThe growing prevalence of health care ethics consultation (HCEC) services in the U.S. has been accompanied by an increase in calls for accountability and quality assurance, and for the debates surrounding why and how HCEC is evaluated. The objective of this study was to evaluate the effectiveness of HCEC as indicated by several novel outcome measurements in East Asian medical encounters.MethodsPatients with medical uncertainty or conflict regarding value-laden issues, and requests made by the attending physicians or nurses for HCEC from December 1, 2009 to April 30, 2012 were randomly assigned to the usual care group (UC group) and the intervention group (HCEC group). The patients in the HCEC group received HCEC conducted by an individual ethics consultant. Data analysis was based on the intention-to-treat principle. Mann–Whitney test and Chi-squared test were used depending on the scale of measurement.ResultsThirty-three patients (53.23%) were randomly assigned to the HCEC group and 29 patients were randomly assigned to the UC group. Among the 33 patients in the HCEC group, two (6.06%) of them ultimately did not receive a HCEC service. Among the 29 patients in the UC group, four (13.79%) of them received a HCEC service. The survival rate at hospital discharge did not differ between the two groups. Patients in the HCEC group showed significant reductions in the entire ICU stay and entire hospital stay. HCEC significantly facilitated achieving the goal of medical care (p < .01). Furthermore, patients in the HCEC group had a shorter ICU stay and shorter hospital stay after the occurrence of medical uncertainty or conflict regarding value-laden issues than those in the UC group.ConclusionsOur findings demonstrated that HCEC were associated with reduced consumption of medical resources as indicated by shorter entire ICU stay, entire hospital stay, and shorter ICU and hospital stay after the occurrence of the medical uncertainty or conflict regarding value-laden issues. This study also showed that HCEC facilitated achieving a consensus regarding the goal of medical care, which conforms to the goal of HCEC.

Hormone changes in men with spinal cord injuries.

The steady state profiles of 63 men with traumatic spinal cord injuries (24 quadriplegics and 39 paraplegics; average age of 31.2 ± 6.8 yr; 18-44 yr) were studied. The average length of post-traumatic period was 6.2 ± 5.0 yr, ranging from 8 months to 20 yr. It was found that all the subjects had normal serum thyroxine, thyrotropin, cortisol, growth hormone and plasma adrenocorticotropic hormone. Seven cases (11.1%) had low serum triiodothyronine and eight cases (12.7%) had low serum testosterone. On the other hand, 17 cases (27.0%) had hyperprolactinemia; 9 cases (14.3%) had elevated serum testosterone level; 6 cases (9.5%) had elevated serum follicle-stimulating hormone; and 4 cases (6.3%) had elevated serum luteinizing hormone. The level of spinal cord injury, injury period and patient age had no correlation with other serum hormone changes except that quadriplegic subjects had lower serum triiodothyronine than the paraplegic, with a mean of 1.42 ± 0.30 v 1.70 ± 0.36 nmol/liter (P < 0.005). Of the eight subjects who had low serum testosterone, none had elevated gonadotropin. There were also eight subjects with elevated follicle-stimulating hormone and/or luteinizing hormone, six of them had normal serum testosterone and two had elevated serum testosterone. This suggested their hypogonadism did not result primarily from classic primary gonadal failure. It could be speculated that other testicular paracrine factors and/or alteration of hypothalamus-pituitarytesticular axis are involved in the pathogenesis of hypogonadism. Further studies in this field will provide information regarding male reproductive physiology and may have impact on fertility enhancement options for men with spinal cord injuries.

Thyroxine Inner Ring Monodeiodinating Activity in Fetal Tissues of the Rat

ABSTRACT: We studied thyroxine (T4) inner ring monodeiodinating activity (5-MA) in various tissues of fetal, maternal, and adult male rats. Tissue homogenates were incubated with 0.26 μM T4 in 0.1 M phosphate buffer (pH 7.4) containing 10 mM EDTA and 400 mM dithiothreitol (final volume 0.7 ml) for 10 min at 37° C; the 3,3‘,5’-triiodothyronine (rT3) generated was measured by radio-immunoassay of ethanol extracts of incubation mixture and the result was corrected for rT3 degradation during incubation. Compared to maternal tissues, T4 to rT3 5-MA in the 14-day-old fetus was increased about 70 times in skeletal muscle (mean ± SEM, velocity, 5.4 ± 0.9 versus 0.08 ± 0.01, pmol rT3/h/mg protein); ∼8 times in intestine (0.72 ± 0.17 versus 0.09 ± 0.03); and ∼4 times in cerebral cortex (19 ± 0.5 versus 4.5 ± 0.9), while it was similar in skin (3.2 ± 0.48 versus 2.6 ± 0.52). Hepatic T4 5-MA approximated 1.1 ± 0.63 in the 14-day-old fetus; it could not be measured reliably in maternal or 19-day fetal tissue because of extensive (>90%) degradation of rT3 during incubation. Relative to mother, T4 5-MA in 19-day fetal tissues was increased ∼30-fold in intestine, ∼20-fold in skeletal muscle, and ∼6-fold in cerebral cortex while it was similar in skin. The T4 5-MA in maternal rat tissues did not differ significantly from corresponding values in adult male rat, except skin, where it was lower in the mother rat (2.6 ± 0.52 versus 4.6 ± 0.61, p < 0.05). In summary, relative to adult tissues T4 5-MA is exceedingly active in several fetal tissues, most notably in skeletal muscle followed by intestine and cerebral cortex.

Impaired Hypothalamus-pituitary-adrenal Axis In Men With Spinal Cord Injuries

Huang T-S, Wang Y-H, Lee S-H, Lai J-S: Impaired hypothalamus-pitutary-adrenal axis in men with spinal cord injuries. Am J Phys Med Rehabil 1998; 77:108–112 Twenty-five men with spinal cord injuries were studied for evaluation of the hypothalamus-pituitary-adrenal axis, using corticotropin-releasing hormone and insulin-induced hypoglycemia. Twenty-five age-matched healthy male volunteers served as controls. Three spinal cord-injured subjects had hyperprolactinemia, three had elevated basal follicle-stimulating hormone levels, one had an elevated basal luteinizing hormone level, and four had hypotestosteronemia. The mean plasma adrenocorticotropin response to corticotropin-releasing hormone of spinal cordinjured subjects was smaller than that of the healthy controls but did not reach a statistical significance. The cortisol response to corticotropin-releasing hormone of the spinal cord-injured subjects was significantly lower than that of healthy controls. However, the difference disappeared if a correction was made for baseline values. Six spinal cord-injured subjects did not have a cortisol response to insulin-induced hypoglycemia, and they had either a minimal or no adrenocorticotropin response. Another 11 spinal cord-injured subjects had a maximal cortisol response to insulin-induced hypoglycemia below the lowest limit of normal, i.e., 0.5 μmol/1. Among these spinal cord-injured subjects, three had a less than 50% increase of plasma adrenocorticotropin after insulin-induced hypoglycemia. These findings are consistent with the notion that spinal cord-injured subjects have an altered central neurotransmitter tone and substantiate the hypothesis that an afferent neural pathway exists between the adrenal and hypothalamus and may modulate stressinduced secretion of adrenocorticotropin. Long-term abnormal adrenocorticotropin secretion may cause mild adrenocortical atrophy and, thereby, a reduced cortisol response.

Synergistic anti-cancer effect of baicalein and silymarin on human hepatoma HepG2 Cells.

This study investigated the effect of baicalein, silymarin, and their combination, on two human liver-derived cell lines, HepG2 (hepatocellular carcinoma) and Chang liver (non-tumor liver cells). It was found that 6.75 microg/ml baicalein or 100 microg/ml silymarin alone significantly inhibited the growth of HepG2. When baicalein was used in combination with silymarin on HepG2, an additive effect at 24 h and a synergistic effect at 48 h were observed. The viability at 48 h was 85.62% from 6.75 microg/ml baicalein treatment; but the viability reduced to 49.67%, 38.56%, and 19.61% when 25, 50, and 100 microg/ml silymarin respectively, was added to the treatment. By contrast, each treatment had little or no effect on Chang liver. Compared to treatment of baicalein or silymarin alone on HepG2, combination of both drugs synergistically increased the percentages of cells in G0/G1 phase and decreased those in S-phase, which were associated with up-regulation of Rb, p53, p21(Cip1) and p27(Kip1) and down-regulation of cyclin D1, cyclin E, CDK4 and phospho-Rb. The results indicate that the combination of baicalein and silymarin eradicates tumor cells efficiently, has minimal deleterious effects to the surrounding normal cells, and offers mechanistic insight for further exploitation of HCC treatment.

Effect of humic acids on thyroidal function

Humic substances (HS) have been implicated as environmental goitrogens. Increased prevalence of goiter has been recently noticed in the blackfoot disease endemic area on the southwest coast of Taiwan, where well water is rich in HS. This study investigated the in vivo effects of humic acids (HA) on the thyroid gland of rats and mice. Groups of mice and rats were fed regular or moderately iodine deficient (∼167 vs 700 μg I− per kg) chow and distilled water or HA water (1 mg/ml) for 3 or 4 months. Serum T4, T3, reverse T3, and/or TSH were measured by radioimmunoassay. Thyroidal 125I uptake was measured in mice at 2 h after injection of 1 μCi125I ip. Treatment of the rat with HA was associated with a significantly (p<0.05) reduced serum T4 without a change in other parameters of study. Treatment with low iodine diet was associated with a clear increase in serum T3 and a decrease in serum rT3. Rats treated with both HA and low iodine diet showed a significantly reduced serum T4, increased serum T3 and decreased serum rT3. In mice, treatment with low iodine diet significantly increased thyroidal 125I uptake and additional treatment with HA significantly enhanced the effect of low iodine diet. Treatment with HA did not influence thyroid weight of rats or mice given normal or iodine deficient diets. We conclude that HA per se do not induce goiter, but they may enhance the goitrogenic effect of low iodine.