Ursolic Acid Induces Apoptotic Cell Death Through AIF and Endo G Release Through a Mitochondria-dependent Pathway in NCI-H292 Human Lung Cancer Cells In Vitro.

BACKGROUND/AIM: Ursolic acid (UA), a triterpene compound present in natural plants, has been shown to induce cytotoxic effects on many human cancer cells through induction of cell-cycle arrest and apoptosis. This study investigated the effects of UA on human lung cancer NCI-H292 cells in vitro. MATERIALS AND METHODS: Flow cytometric assay was used to measure the percentage of cell viability, apoptotic cell death by double staining of annexin V and propidium iodide (PI), production of reactive oxygen species (ROS) and Ca2+, and mitochondriaI membrane potential (Ψm). UA-induced chromatin condensation and DNA fragmentation were examined by 4',6-diamidino-2-phenylindole staining and DNA gel electrophoresis, respectively. Western blotting was used to examine the changes of apoptosis-associated protein expression in NCI-H292 cells. RESULTS: UA reduced cell viability and induced apoptotic cell death. UA increased Ca2+ production, reduced Ψm, but did not affect ROS production in NCI-H292 cells. UA increased apoptosis-inducing factor (AIF) and endonuclease G in NCI-H292 cells. CONCLUSION: Based on these observations, we suggest UA induces apoptotic cell death via AIF and Endo G release through a mitochondria-dependent pathway in NCI-H292 cells.

Bufalin Enhances Immune Responses in Leukemic Mice Through Enhancing Phagocytosis of Macrophage In Vivo.

BACKGROUND/AIM: Bufalin, bufadienolide present in Chan Su, has been shown to induce cancer cell apoptosis in many human cancer cells, including human leukemia cells, but its effects on immune responses are unknown. MATERIALS AND METHODS: This study investigated whether bufalin affected immune responses of mice with WEHI-3 cell-generated leukemia in vivo. BALB/c mice were intraperitoneally injected with WEHI-3 cells to develop leukemia and then were treated with oral treatment with bufalin at different doses (0, 0.1, 0.2 and 0.4 mg/kg) for 2 weeks. At the end of treatment, all mice were weighted and blood was collected; liver and spleen tissues were collected for cell marker, phagocytosis, natural killer (NK) cell activity and T- and B-cell proliferation measurements by using flow cytometric assays. RESULTS: When compared with the leukemia control group, bufalin increased the body weight, but reduced liver and spleen weights, and reduced CD3, CD16 and Mac-3 cell markers at 0.4 mg/kg treatment and increased CD11b marker at 0.1 and 0.2 mg/kg treatment. Furthermore, bufalin at 0.4 mg/kg increased phagocytosis by macrophages isolated from peripheral blood mononuclear cells and at 0.1 mg/kg by those from the peritoneal cavity. Bufalin (0.2 and 0.4 mg/kg) increased NK cell cytotoxic activity at effector:target ratio of 50:1. Bufalin increased B-cell proliferation at 0.1 and 0.2 mg/kg treatment but only increased T-cell proliferation at 0.1 mg/kg. Bufalin increased glutamate oxaloacetate transaminase level at all dose treatments, increased glutamic pyruvic transaminase level only at 0.1 mg/kg treatment, but reduced the level of lactate dehydrogenase at all dose levels in mice with WEHI-3 cell-induced leukemia in vivo. CONCLUSION: Bufalin increased immune responses by enhancing phagocytosis in mice with leukemia mice.

Laminarin Promotes Immune Responses and Reduces Lactate Dehydrogenase But Increases Glutamic Pyruvic Transaminase in Normal Mice In Vivo.

BACKGROUND/AIM: Laminarin, a typical component of fungal cell walls, has been shown to induce immune responses in both adult and larval locusts. We investigated the effects of laminarin on immune response and glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT) and lactate dehydrogenase (LDH) levels in normal mice. MATERIALS AND METHODS: Thirty-six normal BALB/c mice were randomly divided into four groups and treatments were provided by gavage. Group I mice acted as normal control; mice of groups II-IV received laminarin at different doses (100 µl at 1, 2.5 and 5.0 mg/mouse in double-distilled water, respectively). All animals were treated for 14 days and were weighed, blood was collected for determination of cell markers, liver and spleen samples were weighed. Spleens were used for phagocytosis and determination of natural killer (NK) cell activity and cell proliferation by flow cytometric assay. RESULTS: Laminarin reduced the body weights and weights of liver and spleen. Laminarin increased CD3, CD19 and Mac-3 cell populations at 2.5 and 5 mg/mouse, however, these did not affect CD11b marker levels. Laminarin (1 and 5 mg/mouse) reduced macrophage phagocytosis from peripheral blood mononuclear cells, but did not affect phagocytosis by macrophages from the peritoneal cavity. At an effector:target ratio of 50:1, laminarin reduced NK cell cytotoxic activity at all levels, but at a ratio of 25:1, only at 1 mg treatment. Laminarin did not affect T-cell and B-cell proliferation. Laminarin increased the level of GPT and reduced that of LDH at all doses, indicating laminarin can protect against liver injury. Laminarin is worthy of investigation in future experiments on improving immune responses.

Chitosan promotes immune responses, ameliorating total mature white blood cell numbers, but increases glutamic oxaloacetic transaminase and glutamic pyruvic transaminase, and ameliorates lactate dehydrogenase levels in leukemia mice in vivo.

The aim of the present study was to investigate the effect of chitosan (a naturally derived polymer) on the immune responses and glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT) and lactate dehydrogenase (LDH) levels in WEHI­3 cell­generated leukemia mice. Mice were divided into control, WEHI­3 control, acetic acid (vehicle)­treated, and 5 and 20 mg/kg chitosan­treated groups. Mice were subsequently weighed, blood was collected, and liver and spleen samples were isolated and weighed. Blood samples were measured for cell markers, the spleen underwent phagocytosis and natural killer (NK) cell activity examination, and cell proliferation was analyzed by flow cytometry. Chitosan did not significantly affect the weights of body, liver and spleen at 5 and 20 mg/kg treatment. Chitosan increased the percentage of CD3 (T cells marker), decreased the levels of CD19 (B­cell marker) and CD11b at 5 mg/kg treatment, and decreased the levels of Mac­3 at 5 and 20 mg/kg treatment. Chitosan significantly increased macrophage phagocytosis of PBMCs, but did not significantly affect macrophage phagocytosis in the peritoneal cavity. Chitosan treatment did not significantly affect the cytotoxic activity of NK cells, and also did not affect T- and B-cell proliferation. Chitosan significantly increased total white blood cell numbers, and GOT and GPT activities were both significantly increased. However, chitosan did not significantly affect LDH activity in leukemia mice. Chitosan may aid in future studies on improving immune responses in the treatment of leukemia.

Chitosan promotes immune responses, ameliorates glutamic oxaloacetic transaminase and glutamic pyruvic transaminase, but enhances lactate dehydrogenase levels in normal mice in vivo.

Chitosan, a naturally derived polymer, has been shown to possess antimicrobial and anti-inflammatory properties; however, little is known about the effect of chitosan on the immune responses and glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT) and lactate dehydrogenase (LDH) activities in normal mice. The aim of the present study was to investigate whether chitosan has an effect on the immune responses and GOT, GPT and LDH activities in mice in vivo. BALB/c mice were divided into four groups. The negative control group was treated with a normal diet; the positive control group was treated with a normal diet plus orally administered acetic acid and two treatment groups were treated with a normal diet plus orally administered chitosan in acetic acid at doses of 5 and 20 mg/kg, respectively, every other day for 24 days. Mice were weighed during the treatment, and following the treatment, blood was collected, and liver and spleen samples were isolated and weighted. The blood samples were used for measurement of white blood cell markers, and the spleen samples were used for analysis of phagocytosis, natural killer (NK) cell activity and cell proliferation using flow cytometry. The results indicated that chitosan did not markedly affect the body, liver and spleen weights at either dose. Chitosan increased the percentages of CD3 (T-cell marker), CD19 (B-cell marker), CD11b (monocytes) and Mac-3 (macrophages) when compared with the control group. However, chitosan did not affect the phagocytic activity of macrophages in peripheral blood mononuclear cells, although it decreased it in the peritoneal cavity. Treatment with 20 mg/kg chitosan led to a reduction in the cytotoxic activity of NK cells at an effector to target ratio of 25:1. Chitosan did not significantly promote B-cell proliferation in lipopolysaccharide-pretreated cells, but significantly decreased T-cell proliferation in concanavalin A-pretreated cells, and decreased the activity of GOT and GPT compared with that in the acetic acid-treated group,. In addition, it significantly increased LDH activity, to a level similar to that in normal mice, indicating that chitosan can protect against liver injury.

Evaluation of Hirsutella sinensis mycelium on food safety and anti-hepatoma activity in an animal model.

There is evidence that Hirsutella sinensis may have antitumor activity. The aim of the present study was to determine the anti-hepatoma effects and food safety assessment of Hirsutella sinensis mycelium in vivo and in vitro. Effects on mutagenicity were determined using a bacterial reverse mutation assay employing the Salmonella typhimurium strains TA98, TA100, TA102, TA1535 and TA1537. There were no dose-dependent increases or decreases in the number of colonies both with and without metabolic S9 activation in Ames tests. Mice were inoculated with SK-Hep 1 cells and those developing tumors were treated with three different concentrations of Hirsutella sinensis mycelium. After six weeks, blood samples were collected and liver pathology was determined. Aspartate aminotransferase levels were significantly different only in the low-dose treatment group (106±27 IU/l, p=0.048), compared to the control group (162±80 IU/l). The tumor weight was significantly different only in the low-dose treatment group. We found that necrosis, hemorrhage and calcifications were presented in both control and experimental groups. Inhibition of tumor growth was observed only at the lowest dose.

Renal calcification in very low birth weight infants.

BACKGROUND: Renal calcification in preterm infants has been described frequently. The etiologic factors have not yet been fully clarified. The objective of this study was to evaluate the incidence of and risk factors for renal calcification in our population. METHODS: We retrospectively reviewed the charts of very low birth weight preterm infants during a 1-year period. Renal ultrasound scans were performed at term or before discharge and at a corrected age of 1 year. RESULTS: Six infants (6%) had renal calcification at term or before discharge compared with 96 who did not. Factors significantly associated with renal calcification included gestational age (26 weeks vs. 29 weeks, p=0.006), birth weight (851 g vs. 1141 g, p=0.004), duration of mechanical ventilation (69 days vs. 29 days, p=0.002), length of intensive care (72 days vs. 41 days, p=0.013), furosemide therapy (33% vs. 3%, p=0.027), and dexamethasone therapy (50% vs. 2% p=0.001). Birth weight and dexamethasone therapy had significant independent association after stepwise logistic regression analysis. Sex, oliguria, acidosis, duration of oxygen therapy, length of hospital stay, nutrition status, and nephrotoxic drugs did not differ between the two groups. Three of the six infants had spontaneous remission of renal calcification, whereas two patients without the finding in neonatal stage had renal calcification at a corrected age of 1 year. CONCLUSION: The incidence of renal calcification in very low birth weight infants in this study was relatively low, and the calcification was transient in one-half of the infants. Extremely premature, sick infants requiring long-term ventilation, and those receiving furosemide or dexamethasone were more likely to have renal calcification. Clinicians should be aware that renal calcification may develop beyond the neonatal stage.

Transcutaneous electrical stimulation of acupoints changes body composition and heart rate variability in postmenopausal women with obesity.

This study aimed to evaluate the effect of transcutaneous electric acupoint stimulations (TEAS) on body composition and heart rate variability (HRV) in postmenopausal women with obesity. In this prospective study, 49 postmenopausal women were recruited in Taiwan. Body composition was used as a screening test for obesity (percentage body fat > 30%, waist circumference > 80 cm). The experimental group (n = 24) received TEAS treatment 30 min twice per week for 12 weeks at the Zusanli (ST 36) and Sanyinjiao (SP 6) acupoints. The control group (n = 25) did not receive any intervention. The study of HRV was analyzed by time (standard deviation of the normal-to-normal (NN) intervals (SDNN) and square root of the mean squared differences of successive NN intervals (RMSSD) indices) and frequency domain methods. Power spectral components were obtained at low (LF) and high (HF) frequencies. Body composition and HRV values were measured at the 4th, 8th, and 12th weeks. A total of 40 subjects completed this study. Waist circumference and percentage body fat in the experimental group (n = 20) were significantly less than those of the control group (n = 20) at the 8th and 12th weeks (all P < .05). Additionally, at the same time points, percentage lean body mass in the experimental group was significantly greater than that in the control group (P < .05). SDNN values increased significantly at the 4th and 8th weeks when compared with the control group (all P < .05). At 12 weeks, SDNN value was not significantly different from that of the control group (P = .105). TEAS treatment improves body composition, and has a transient effect on the HRV in postmenopausal women with obesity.

Effects of acupuncture-like transcutaneous electrical nerve stimulation on children with asthma.

OBJECTIVE: To evaluate the effects of acupuncture-like transcutaneous electrical nerve stimulation (AL-TENS) on children with asthma. METHODS: After an 8-week run-in period, the experimental group were assigned to treatment with AL-TENS, whereas the control group did not receive AL-TENS. A total of 43 children with asthma were recruited from a hospital and an elementary school. All the cases had been diagnosed as having asthma by physicians. The outcome measures included pulmonary function tests (PFTs), heart rate turbulence (HRT), heart rate variability (HRV), and pediatric asthma quality-of-life questionnaire (PAQLQ). RESULTS: After 8 weeks of AL-TENS, there were no significant differences on forced vital capacity (FVC), FEV1/FVC, and peak expiratory flows (PEFs) between the two groups. The HRT is the physiological, biphasic response of the sinus node to premature ventricular contractions. In the experimental group, the mean HRT was statistically significant between pretest and posttest. The HRV and the PAQLQ showed no difference, but in the experimental group, the subscale of the PAQLQ (particularly activity) improved significantly more than the pretest scores. Furthermore, there were no differences in PFTs and HRV after 8 weeks of AL-TENS between the two groups. CONCLUSIONS: The PAQLQ activities of the experimental group improved significantly more than those of the control group. Reasons may include: 1) the asthma cases were stable and the cases were on stable status and 2) the degree of airway remodeling was less. It is suggested that in the future, treatment frequency and the long-term follow-up for evaluating the effects of AL-TENS on children with asthma should also be considered.